Study-unit IMAGING TECHNOLOGIES

Course name Biotechnology
Study-unit Code A000992
Curriculum Comune a tutti i curricula
Lecturer Maurizio Mattarelli
Lecturers
  • Maurizio Mattarelli
Hours
  • 42 ore - Maurizio Mattarelli
CFU 6
Course Regulation Coorte 2022
Supplied 2024/25
Supplied other course regulation
Learning activities Affine/integrativa
Area Attività formative affini o integrative
Sector FIS/03
Type of study-unit Opzionale (Optional)
Type of learning activities Attività formativa monodisciplinare
Language of instruction Italian
Contents
Elements of electromagnetism, wave optics, and geometric optics for understanding imaging techniques.
Operating principle and application examples of the main microscopy techniques, relevant for biotechnological applications.

Reference texts
Fundamentals of Light Microscopy and Electronic Imaging, 2nd Edition Douglas B. Murphy Michael W. Davidson (Wiley-Blackwell Ed). Also available in the electronic resources of the University (in particular on Proquest Ebook Central).

A university text of General Physics II (electromagnetism and optics).
For example: P. Mazzoldi, M. Nigro, C. Voices, Elements of Physics, Vol. 2, (Ed. EdiSES).
D. Halliday, R. Resnik, J. Walker, Fundamentals of Physics (Vol. 2), Ambrosiana Publishing House, Milan.

Lecture notes, articles and textbook chapters indicated therein (available in the University's online resources).

Educational objectives
The main objective of the course is to provide students with the basic knowledge to understand the theoretical foundation and some experimental method of bio-imaging.
The main knowledge acquired will concern the principles and potential of some experimental methods of optical and spectroscopic imaging, electron and scanning probe microscopy .
The main skills that the course proposes to transmit are:
- know how to use the fundamental physical laws to understand the operating principle of modern imaging technologies, their potential and their limits.
- know how to use and analyze the results of the main imaging methods for relevant applications in the field of Biotechnology.

Prerequisites
In order to understand the contents of the Study-unit, it is advisable to have acquired basic knowledge of Physics, such as those provided in the Study-unit Physics in the first year of the Biotechnology course.

Teaching methods
Lectures are provided on all the topics covered by the course and the analysis of experiments concerning optical microscopies, SEM, and spectroscopic imaging.
Learning verification modality
The assessment of the course's educational objectives involves an oral exam lasting approximately 30 minutes. The exam will consist of a brief presentation of an imaging technique studied during the course (approximately 15 minutes) based on an experimental case in the biological field taken from a scientific article.

The presentation should include a description of the instrumentation, the physical interaction mechanism, and the technique's resolution capabilities. In addition to possible requests for clarification or further detail during the presentation, the exam will then continue with one or two questions aimed at assessing the breadth of the student's preparation.

The purpose of the exam is to verify: 1) the knowledge acquired regarding theoretical issues and the various techniques and instruments studied during the course; 2) the acquisition of critical skills aimed at choosing the most suitable experimental technique for studying the sample in question; 3) the ability to communicate effectively and relevantly in oral form.
The final evaluation will be determined by the Commission on a scale of thirty.

For information on support services for students with disabilities and/or learning disorders, visit the page http://www.unipg.it/disabilita-e-dsa
Extended program
Electromagnetic fields and waves. Basics on the polarization of matter. Refractive index. Absorption.

Elements of geometric optics. Refraction and Reflection. Snell's Laws. Limit angle. Lenses and mirrors. Optical systems: eye, telescope, microscope. Optical aberration.

Coherent Waves. Young's experiment. Interference and diffraction. Airy disk and diffraction limit. Interference from thin films. Bragg's law.
Spectral analysis. Diffraction grating. Interferometer. Optical filters. Spectral resolution.

Photoelectric effect and quantum nature of radiation.
Light sources: Sun, incandescent lamps, gas lamps, LEDs. LASER.
Thermal detectors. Photomultipliers. Detector a semiconductor. Photodiodes. Spatial detectors (FPA, CCD, CMOS).

Hyperspectral reflectance imaging. Color: stimulus and perception. Diffuse and specular reflectance. Colorimetry. The human eye. Mapping strategies. Application to the study of vegetation. Devices. Spectral indices.

Optical microscopy. Elements of a combined microscope. Illumination. Lateral and axial spatial resolution. Abbe's limit. Rayleigh criterion. Contrast formation. Microscopy: bright field, dark field, phase contrast, polarized light. Fluorescence microscopy. Physical mechanism of fluorescence. Fluorescent molecules. Markers. Techniques: FRET, FRAP, FLIM. Confocal fluorescence microscopy.
Strategies for increasing the resolution: engineering of the scattering volume and non-linear excitation. Techniques: 4Pi, SIM, SMLM STED, TIRF, 2P, SNOM.

Vibrational Spectroscopy. Normal modes of vibration in molecules and solids. Infrared absorption. FTIR microscopy. Raman effect. Raman microscopy. Measuring equipment. Basics of Brillouin spectroscopy. Combined optical systems.

Electron microscopy. Wavelength of electrons. Instrumentation: sources, lenses, detectors. Electron-matter interaction. Secondary and backscattered electrons. EDX. Electronic spectroscopy. Spatial resolution. Systems: SEM, TEM, STEM. Preparation of biological samples for electron microscopy.
Scanning probe microscopy. Scanning tunneling microscope. Atomic force microscopy. Imaging Modes: contact, tapping, non contact. AFM of biological samples.

Digital images. Sampling. Nyquist criterion. Characteristics of detectors. Image processing. Calibration. Digital filters. Segmentation. Dimensional reduction of multispectral images.